慢性束缚应激对小鼠睾丸生精功能及m6A甲基化酶表达的影响  

Effects of chronic restraint stress on spermatogenesis and m6A methylase expression in mouse testis

作　　者：孟星圻 彭莉轩 唐筱涵 易康 张巳 向宇燕 李素云 Meng Xingqi;Peng Lixuan;Tang Xiaohan;Yi Kang;Zhang Si;Xiang Yuyan;Li Suyun(Clinical Anatomy&Reproductive Medicine Aplication Insititute,Hengyang Medical College,University of South China,Hengyang 421001,Hunan,Chin)

机构地区：[1]南华大学衡阳医学院应用解剖学与生殖医学研究所,湖南衡阳421001

出　　处：《中国男科学杂志》2023年第3期19-25,共7页Chinese Journal of Andrology

基　　金：湖南省自然科学基金项目(No.2021JJ30593,2021JJ30598);湖南省大学生创新创业训练计划项目(No.S202110555300,S202110555316);湖南省教育厅基金项目(No.21C0301)。

摘　　要：目的探讨慢性束缚应激(CRS)对小鼠睾丸生精功能及m6A甲基化酶的影响。方法18只8周龄雄性C57 BL/6小鼠,随机分为空白对照组(Control)和慢性束缚应激组(CRS)(每天在圆筒内慢性束缚6 h,连续35 d)。小鼠睾丸和附睾尾称重,进行精子计数,精子活率、畸形率检测及对睾丸形态结构变化等进行H&E染色检测;使用qRT-PCR和Western blot检测各组小鼠睾丸组织中m6A甲基化酶mRNA及蛋白表达的变化。结果与Control组相比,CRS组小鼠睾丸以及附睾重量有下降的趋势,精子数量下降、精子畸形率增高及精子活率降低差异有统计学意义;与Control组相比,CRS组小鼠睾丸形态结构破坏,各级生精细胞排列紊乱,管腔内精子数量减少;qRT-PCR结果显示,与Control组相比,CRS组小鼠睾丸m6A甲基化转移酶WTAP mRNA表达降低,去甲基化酶ALKBH5及甲基识别酶YTHDC1、YTHDC2、YTHDF1、YTHDF3 mRNA表达增高,差异有统计学意义。Western blot结果显示,与Control组相比,CRS组小鼠睾丸m6A甲基化转移酶WTAP蛋白表达降低,去甲基化酶ALKBH5及甲基识别酶YTHDC1蛋白表达增高,差异有统计学意义。结论CRS导致小鼠出现生精功能障碍,其生精障碍可能与睾丸内m6A甲基化酶异常变化有关。Objective To investigate the effects of chronic restraint stress(CRS)on spermatogenesis and m6A methylase expression in mouse testis.Methods Eighteen 8⁃week⁃old male C57 BL/6 mice were randomly divided into blank control group(Control)and chronic restraint stress group(CRS).The weight of testis and epididymis tail was weighed,the sperm count of mice was measured,and the sperm activity,deformity rate and testicular morphology were observed by HE staining;mRNA and protein expression of m6A methylase in testicular tissue of mice in each group was detected by qRT⁃PCR and Western blot,respectively.Results Compared with that in the control group,the weight of testis and epididymis in the CRS group showed a downward trend.Simultaneously,an obvious decrease in the number and viability of sperm and an significant increase in sperm deformity rate were found;In addition,mRNA expression of m6A methyltransferase WTAP was noticeably decreased in testis tissues in the CRS group,while the mRNA expressions of demethylase ALKBH5 and methyl recognition enzymes YTHDC1,YTHDC2,YTHDF1,YTHDF3 were remarkably increased.Similarly,the protein expression levels of the above indexes were consistent with their mRNA expression levels.Conclusion CRS⁃induced spermatogenesis disorder in mice may be related to abnormal expression of m6A methylase in testis.

关 键 词：慢性束缚应激 小鼠 生精功能障碍 m6A甲基化酶 

分 类 号：R339.211[医药卫生—人体生理学]